Abstract:
The ex vivo myofiber culture system has proven to be a useful methodology to explore the biology and behavior of satellite cells within their niche environment. However, a limitation of this system is that myofibers and their associated satellite cells are commonly examined using conventional fluorescence microscopy, which renders a three-dimensional system into two-dimensional imaging, leading to the loss of precious information or misleading interpretation of observations. Here, we report on the use of light-sheet fluorescence microscopy to generate three-dimensional and live imaging of satellite cells on myofibers. Light-sheet microscopy offers high imaging speed and good spatial resolution with minimal photo-bleaching, allowing live imaging and three-dimensional acquisition of skeletal muscle fiber specimen. The potentials of this technology are wide, ranging from the visualization of satellite cell behavior such as cell division and cell migration to imaging the sub-cellular localization of proteins or organelles.
摘要:
离体肌纤维培养系统已被证明是探索卫星细胞在其生态位环境中的生物学和行为的有用方法。然而,该系统的局限性在于,肌纤维及其相关的卫星细胞通常使用常规的荧光显微镜检查,将三维系统转化为二维成像,导致宝贵信息的丢失或对观察结果的误导性解释。这里,我们报告了使用光片荧光显微镜对肌纤维上的卫星细胞进行三维和活体成像。光片显微镜提供高成像速度和良好的空间分辨率与最小的光漂白,允许实时成像和骨骼肌纤维标本的三维采集。这项技术的潜力很大,从卫星细胞行为(如细胞分裂和细胞迁移)的可视化到蛋白质或细胞器的亚细胞定位成像。